The infectious cDNA clones of HEV G1 (pSK-HEV-2 from the Sar-55 strain) (15 (link)) and G3 (p6 from the Kernow-C1 strain) were generous gifts from Dr. Suzanne U. Emerson (NIH, Bethesda, MD), whereas the infectious cDNA clone of G4 (pHEV-4TW from the TW6196E strain) has been described previously (37 (link)). The Huh7-S10-3 cell line was also a gift from Dr. Emerson. S10-3 cells were maintained in Dulbecco's Modified Eagle Medium (DMEM) containing 10% FBS (Gibco), penicillin (250 IU/mL) and streptomycin (250 μg/mL) at 37°C in the presence of 5% CO2. MRT67307, BX795, and ribavirin were purchased from Selleck Chemicals, and peg-IFNα-2a was purchased from Shanghai Roche Pharmaceuticals.
The G3 Kernow-C1 HEV infectious virus stock used for gerbil inoculation was rescued from the p6 infectious cDNA clones in S10-3 cells as described previously (16 (link), 36 (link)). The viral RNA titer of the rescued infectious HEV stock was determined by real-time qRT-PCR (56 (link)), with 6.83 × 106 genome equivalents (GE) of viral RNA/100 μL medium, giving an infectivity of 1 fluorescent focus-forming unit (FFU)/5,618 GE, as determined previously (50 (link)).
The G3 Kernow-C1 HEV infectious virus stock used for gerbil inoculation was rescued from the p6 infectious cDNA clones in S10-3 cells as described previously (16 (link), 36 (link)). The viral RNA titer of the rescued infectious HEV stock was determined by real-time qRT-PCR (56 (link)), with 6.83 × 106 genome equivalents (GE) of viral RNA/100 μL medium, giving an infectivity of 1 fluorescent focus-forming unit (FFU)/5,618 GE, as determined previously (50 (link)).
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